A method for rapid reconstitution of ADP/ATP carrier from Jerusalem artichoke (Helianthus tuberosus L.) tubers mitochondria in proteoliposomes is described. The method is based on the well known property of the Amberlite resin to absorb the detergent allowing proteoliposome formation. This has been achieved by a micro-batchwise technique, using a rotating plate stirrer. An evaluation of the optimal conditions, in comparison with the more usual column method is presented. The purified ADP/ATP carrier, incorporated in proteoliposomes by this method, shows a high transport activity and a higher specific activity with respect to proteoliposomes obtained by the column procedure. Furthermore the proteoliposomal preparations are more homogeneous in size, with a diameter ranging from 300 to 350 nm. The method is suitable for the reconstitution of other membrane transport proteins.
A micro-bachwise technique method for rapid reconstitution of functionally active mitochondrial ADP/ATP carrier from Jerusalem artichoke (Helianthus tuberosus L.) tubers
DE PALMA, Annalisa;SCALERA, Vito Dom. E.
2008-01-01
Abstract
A method for rapid reconstitution of ADP/ATP carrier from Jerusalem artichoke (Helianthus tuberosus L.) tubers mitochondria in proteoliposomes is described. The method is based on the well known property of the Amberlite resin to absorb the detergent allowing proteoliposome formation. This has been achieved by a micro-batchwise technique, using a rotating plate stirrer. An evaluation of the optimal conditions, in comparison with the more usual column method is presented. The purified ADP/ATP carrier, incorporated in proteoliposomes by this method, shows a high transport activity and a higher specific activity with respect to proteoliposomes obtained by the column procedure. Furthermore the proteoliposomal preparations are more homogeneous in size, with a diameter ranging from 300 to 350 nm. The method is suitable for the reconstitution of other membrane transport proteins.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
